Core engagement with β-arrestin is dispensable for agonist-induced vasopressin receptor endocytosis and ERK activation

Kumari, Punita; Srivastava, Ashish; Ghosh, Eshan; Ranjan, Ravi; Dogra, Shalini; Yadav, Prem N.; Shukla, Arun K.

doi:10.1091/mbc.e16-12-0818

Cited by 97 publications

(93 citation statements)

References 40 publications

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“…Active conformation of the receptor and subsequent GRK-mediated phosphorylation are generally thought to be prerequisite for β-arrestin binding (16), suggesting that the two main interactions occur simultaneously. Recent studies have shown that the phosphate and the core interactions can be separated in vitro or by using receptor chimeras and mutants (51)(52)(53). However, the question remained whether the distinct interactions could be independent in physiologically relevant situations.…”

Section: Discussionmentioning

confidence: 99%

Heterologous phosphorylation–induced formation of a stability lock permits regulation of inactive receptors by β-arrestins

Tóth

Prokop

Gyombolai

et al. 2018

Journal of Biological Chemistry

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β-arrestins are key regulators and signal transducers of G protein-coupled receptors (GPCRs). The interaction between receptorsand β-arrestins is generally believed to require both receptor activity and phosphorylation by GPCR kinases. In this study, we investigated whether β-arrestins are able to bind second messenger kinase-phosphorylated, but inactive receptors as well. Since heterologous phosphorylation is a common phenomenon among GPCRs, this mode of β-arrestin activation may represent a novel mechanism of signal transduction and receptor cross-talk.Here we demonstrate that activation of protein kinase C (PKC) by phorbol myristate acetate, G q/11-coupled GPCR or epidermal growth factor receptor stimulation promotes β-arrestin2 recruitment to unliganded AT1 angiotensin receptor (AT1R). We found that this interaction depends on the stability lock, a structure responsible for the sustained binding between GPCRs and β-arrestins, formed by phosphorylated serine-threonine clusters in the receptor's C-terminus and two conserved phosphate-binding lysines in the β-arrestin2 Ndomain. Using improved FlAsH-based β-arrestin2 conformational biosensors, we also show that the stability lock not only stabilizes the receptor-β-arrestin interaction, but also governs the structural rearrangements within β-arrestins. Furthermore, we found that β-arrestin2 binds to PKC-phosphorylated AT1R in a distinct active conformation, which triggers MAPK recruitment and receptor internalization. Our results provide new insights into the activation of β-arrestins and reveal their novel role in receptor cross-talk.The family of G protein-coupled receptors (GPCRs) consists of ~800 members in humans and about 30% of modern drugs target these molecules (1). GPCRs respond to a wide variety of endogenous ligands, including hormones, neurotransmitters, and lipids. Despite their huge diversity, the signal transduction mechanisms of GPCRs share several common features: agonist binding is followed by the activation of a relatively small number of heterotrimeric G proteins, which initiate complex intracellular signaling cascades. Receptor responsiveness to further stimulation is attenuated by a multistep process, called desensitization (2). In case of homologous desensitization, active GPCRs are phosphorylated by GPCR kinases (GRKs) followed by the recruitment of β-arrestin proteins (β-arrestin1 and JBC C o n f i d e n t i a lHeterologous regulation of inactive receptors via β-arrestin 2 β-arrestin2, also known as arrestin-2 and arrestin-3, respectively). β-arrestins uncouple the receptors from G proteins and initiate receptor internalization (3), thereby serving as the key regulators of GPCRs' function. In contrast, heterologous desensitization is mediated by second-messenger activated kinases, such as protein kinase C (PKC), which can phosphorylate active and inactive receptors. Heterologous desensitization was originally thought to be independent of β-arrestins, however, some data have challenged this concept (4-6). In addition to their rol...

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Section: Discussionmentioning

confidence: 99%

Heterologous phosphorylation–induced formation of a stability lock permits regulation of inactive receptors by β-arrestins

Tóth

Prokop

Gyombolai

et al. 2018

Journal of Biological Chemistry

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“…All of the above findings fit with an emerging model whereby GPCRs affect multiple modes of β-arrestin function through interactions at functionally distinct sites. 93,94 Notably, the dependence of ERK1/2 activation on lengthened endocytic lifetimes has so far been demonstrated only with receptors whose activation of MAPK is dependent on β-arrestin.…”

Section: Signaling Consequences Of Receptor-regulated Endocytosismentioning

confidence: 99%

Regulation of G protein‐coupled receptor signaling by plasma membrane organization and endocytosis

Weinberg

Puthenveedu

2019

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The trafficking of G protein coupled‐receptors (GPCRs) is one of the most exciting areas in cell biology because of recent advances demonstrating that GPCR signaling is spatially encoded. GPCRs, acting in a diverse array of physiological systems, can have differential signaling consequences depending on their subcellular localization. At the plasma membrane, GPCR organization could fine‐tune the initial stages of receptor signaling by determining the magnitude of signaling and the type of effectors to which receptors can couple. This organization is mediated by the lipid composition of the plasma membrane, receptor‐receptor interactions, and receptor interactions with intracellular scaffolding proteins. GPCR organization is subsequently changed by ligand binding and the regulated endocytosis of these receptors. Activated GPCRs can modulate the dynamics of their own endocytosis through changing clathrin‐coated pit dynamics, and through the scaffolding adaptor protein β‐arrestin. This endocytic regulation has signaling consequences, predominantly through modulation of the MAPK cascade. This review explores what is known about receptor sorting at the plasma membrane, protein partners that control receptor endocytosis, and the ways in which receptor sorting at the plasma membrane regulates downstream trafficking and signaling.

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“…core-engaged) receptor-βarr complexes, respectively. As recent studies have revealed distinct functional outcomes associated with these two conformations of receptor-βarr complexes (Cahill et al, 2017; Kumari et al, 2016; Kumari et al, 2017; Sente et al, 2018), we envisioned that key determinants of the functional divergence of βarr isoforms may lie at the level of structural and conformational differences between receptor-bound βarrs.…”

Section: Introductionmentioning

confidence: 99%

Conformational sensors and domain-swapping reveal structural and functional differences between β-arrestin isoforms

Ghosh

Dwivedi

Baidya

et al. 2019

Preprint

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SummaryDesensitization, signaling and trafficking of G protein-coupled receptors (GPCRs) are critically regulated by multifunctional adaptor proteins, β-arrestins (βarrs). The two isoforms of βarrs (βarr1 and 2) share a high degree of sequence and structural similarity, still however, they often mediate distinct functional outcomes in the context of GPCR signaling and regulation. A mechanistic basis for such a functional divergence of βarr isoforms is still lacking. Using a set of complementary approaches including antibody fragment based conformational sensors, we discover structural differences between βarr1 and 2 upon their interaction with activated and phosphorylated receptors. Interestingly, domain swapped chimeras of βarrs display robust complementation in functional assays thereby, linking the structural differences between the receptor-bound βarr1 and 2 with their divergent functional outcomes. Our findings reveal important insights into the ability of βarr isoforms to drive distinct functional outcomes, and underscore the importance of integrating this aspect in the current framework of biased agonism.

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Core engagement with β-arrestin is dispensable for agonist-induced vasopressin receptor endocytosis and ERK activation

Cited by 97 publications

References 40 publications

Heterologous phosphorylation–induced formation of a stability lock permits regulation of inactive receptors by β-arrestins

Heterologous phosphorylation–induced formation of a stability lock permits regulation of inactive receptors by β-arrestins

Regulation of G protein‐coupled receptor signaling by plasma membrane organization and endocytosis

Conformational sensors and domain-swapping reveal structural and functional differences between β-arrestin isoforms

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